Long-term cultivation of multipotential neural stem cells from adult rat subependyma

Gobbel, Glenn T.; Choi, Seungjin; Beier, Steven; Niranjan, Ajay

doi:10.1016/s0006-8993(03)02977-9

Cited by 32 publications

(14 citation statements)

References 40 publications

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“…NSC could be successfully isolated, cultured and differentiated using a standard protocol (Gobbel et al, 2003). The NSC were subjected to three differentiation protocols which were specifically designed for neuronal differentiation of bone marrow-derived stem cells.…”

Section: Discussionmentioning

confidence: 99%

“…Neural stem cells were isolated according to a published protocol (Gobbel et al, 2003). In brief, the brain of female Sprague-Dawley rats (n = 2, body weight 250 g, Harlan) was removed and placed in artificial cerebrospinal fluid (124 mM NaCl, 5 mM KCl, 3.2 mM MgCl 2 , 105 mM NaHCO 3 , 10 mM glucose and 2 mM CaCl 2 ).…”

Section: Culture Of Neural Stem Cellsmentioning

confidence: 99%

“…Normalized mRNA expression levels of neuroglial genes in MAPC-like cells and NSC before (respectively M0 and N0) and after differentiation according to the Jiang (M1, N1), the Kohyama (M2, N2) and the Kabos (M3, N3) protocol were compared with levels of expression in an NSC clone that was differentiated following a standard protocol (N standard ). According to this protocol, cells are plated on poly-lysine in medium devoid of growth factors but with the addition of 3% Fetal Bovine Serum (Reynolds and Weiss, 1996;Gobbel et al, 2003). Therefore, the formula to quantify expression of the genes of interest (Genes int ) is: [Relative expression (Gene int …”

Section: Neural Differentiation Of Mapc-like Cells and Nscmentioning

confidence: 99%

“…Neural stem cells were isolated from the subventricular zone of the brain by plating dissociated, subventricular cells in a special defined neural stem cell expansion medium (Gobbel et al, 2003). Two weeks after initial isolation, large spherical clusters of cells, or neurospheres, which grew loosely attached to the substrate, were present in the culture.…”

Section: Isolation and Phenotypic Characterization Of Nsc Cellsmentioning

confidence: 99%

“…Approximately, 4% of the cells formed neurospheres which could be passaged continuously (> 10 passages). Using the culture and isolation technique especially designed for adult rat neural stem cells, we were able to keep single cell clones of NSC in culture for over 1 year (Gobbel et al, 2003). Formation of neurospheres and expansion capacity were used as criteria for the selection of a clone which was used in this study.…”

Section: Isolation and Phenotypic Characterization Of Nsc Cellsmentioning

confidence: 99%

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Differentiation assays of bone marrow-derived Multipotent Adult Progenitor Cell (MAPC)-like cells towards neural cells cannot depend on morphology and a limited set of neural markers

Raedt

Pinxteren

Dycke

et al. 2007

Experimental Neurology

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There are accumulating studies that report a neurogenic potential of bone marrow-derived cells both in vitro as well as in vivo. Most claims of neural "transdifferentiation" have based their conclusions on morphology and neural gene expression. Recently, doubts have been raised about the validity of both outcome parameters since non-neural cells can extend neurites and show aberrant neural gene expression as a response to stress inducing factors. In this study, we compared bone marrow-derived Multipotent Adult Progenitor Cell (MAPC)-like cells and neural stem cells (NSC) in their morphology and neural gene expression profile after neural differentiation using three differentiation protocols. We evaluated the expression of five neuroglial antigens [neurofilament 200 (NF200); beta III tubulin (β3 tub); tau; Glial Fibrillary Acidic Protein (GFAP); Myelin Basic Protein (MBP) and RIP antigen] using real-time PCR (RT-PCR) and immunocytochemistry (ICC). MAPC-like cells adopted a neural-like morphology in one protocol but a fibroblast-like morphology in the two other protocols. RT-PCR and ICC show that MAPC-like cells already express the neural antigens beta III tubulin and NF200 at baseline, but no upregulation of these genes after exposure to three distinct differentiation protocols was seen. In contrast, NSC adopt neural and glial morphologies with a clear increase in expression of all neuroglial genes in all differentiation protocols used. In conclusion, our data demonstrate that neural-like morphology and expression of a limited set of neural marker genes by MAPC-like cells after differentiation are not absolute proof of neural transdifferentiation because MAPC-like cells only partially meet the criteria which are fulfilled by NSC after neural differentiation.

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Section: Discussionmentioning

confidence: 99%

Section: Culture Of Neural Stem Cellsmentioning

confidence: 99%

Section: Neural Differentiation Of Mapc-like Cells and Nscmentioning

confidence: 99%

Section: Isolation and Phenotypic Characterization Of Nsc Cellsmentioning

confidence: 99%

Section: Isolation and Phenotypic Characterization Of Nsc Cellsmentioning

confidence: 99%

See 3 more Smart Citations

Differentiation assays of bone marrow-derived Multipotent Adult Progenitor Cell (MAPC)-like cells towards neural cells cannot depend on morphology and a limited set of neural markers

Raedt

Pinxteren

Dycke

et al. 2007

Experimental Neurology

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Culture of Neural Stem Cells in Calcium Alginate Beads

Li¹,

Liu²,

Song³

et al. 2006

Biotechnol Progress

121

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Neural stem cells (NSCs) with the capacity of extensive self-renewal and multilineage differentiation have attracted more and more attention in research as NSCs will play an important role in the nerve disease treatment and nerve injury repair. The shortage of NSCs, both their sources and their numbers, however, is the biggest challenge for their clinic application, and hence, in vitro culture and expansion of NSCs is vitally important to realize their potentials. In this work, mouse-derived NSCs were cultured in three-dimensional calcium alginate beads (Ca-Alg-Bs). Gelling conditions, cell density, and cell harvest were determined by the exploration of formation and dissociation parameters for Ca-Alg-Bs. Additionally, the recovered and the subsequent induced cells were identified by immunofluorescence staining of Nestin, beta-tubulin, and GFAP. The results show that the 2-mm diameter Ca-Alg-Bs, prepared with 1.5% sodium alginate solution and 3.5% CaCl2 solution and with gelling for 10 min, is suitable for the NSCs culture. The seeding density of 0.8 x 10(5) cells x mL-1 for the encapsulation of NSCs resulted in the most expansion, and the NSCs almost doubled during the experiment. The average cell recovery rate is over 88.5%, with the Ca-Alg-Bs dissolving in 55 mM sodium citrate solution for 10 min. The recovered cells cultured in the Ca-Alg-Bs still expressed Nestin and had the capacity of multilineage differentiation into neurons and glial cells and, thus, remained to be NSCs. These results demonstrate that NSC expansion within Ca-Alg-Bs is feasible and provides further possibilities for NSC expansion in bioreactors of the scale of clinical relevance.

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Functional immobilization of interferon‐gamma induces neuronal differentiation of neural stem cells

Leipzig

Zahir

et al. 2009

J Biomedical Materials Res

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Stem cell transplantation provides significant promise to regenerative strategies after injury in the central nervous system. Neural stem/progenitor cells (NSPCs) have been studied in terms of their regenerative capacity and their ability to differentiate into neurons when exposed to various soluble factors. In this study, interferon-gamma (IFN-gamma) was compared with brain-derived neurotrophic factor (BDNF) and erythropoietin and was shown to be the best single growth factor for inducing neuronal differentiation from adult rat brain-derived NSPCs. Next, IFN-gamma was surface immobilized to a methacrylamide chitosan (MAC) scaffold that was specifically designed to match the modulus of brain tissue and neuronal differentiation of NSPCs was examined in vitro by immunohistochemistry. Bioactive IFN-gamma was successfully immobilized and quantified by ELISA. Both soluble and immobilized IFN-gamma on MAC surfaces showed dose dependent neuronal differentiation with soluble saturation occurring at 100 ng/mL and the most effective immobilized IFN-gamma dose at 37.5 ng/cm(2), where significantly more neurons resulted compared with controls including soluble IFN-gamma.

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Long-term cultivation of multipotential neural stem cells from adult rat subependyma

Cited by 32 publications

References 40 publications

Differentiation assays of bone marrow-derived Multipotent Adult Progenitor Cell (MAPC)-like cells towards neural cells cannot depend on morphology and a limited set of neural markers

Differentiation assays of bone marrow-derived Multipotent Adult Progenitor Cell (MAPC)-like cells towards neural cells cannot depend on morphology and a limited set of neural markers

Culture of Neural Stem Cells in Calcium Alginate Beads

Functional immobilization of interferon‐gamma induces neuronal differentiation of neural stem cells

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